The virotype BVDV RT-PCR Kit allows the safe detection of RNA from bovine viral diarrhea virus genotypes 1 and 2 (BVDV-1 and BVDV-2) in samples from cattle. The kit contains all of the reagents for real-time RT-PCR. It can be used with pools of up to 50 individual samples of blood, plasma, or serum, 25 individual samples of ear tissue, and 100 individual samples of milk.

In Germany: Registered in accordance with §17c of the German Law on Animal Diseases. Registration No.: FLI-B 451

In Switzerland: Approved by the Federal Veterinary Office (BVET)

Performance

virotype BVDV RT-PCR Kit detects RNA from bovine viral diarrhea virus genotypes in samples. A validation study of 5056 samples (EDTA blood, serum, plasma, ear tissue, and milk; kindly provided by state laboratories) with known BVDV status was tested with the virotype BVDV RT-PCR Kit. Of 4570 negative samples, 4568 samples tested negative, providing a diagnostic specificity with this sample set of 99.95%. Of 486 positive samples, 484 samples tested positive, providing a diagnostic sensitivity with this sample set of 99.6% (see the table Specificity and sensitivity of virotype BVDV RT-PCR Kit). In addition, samples from the German BVD ring trial were tested with the virotype BVDV RT-PCR Kit (conducted by the Friedrich-Loeffler-Institut). The sample set contained positive, weak positive, and negative serum, as well as milk and ear tissue samples. The virotype BVDV RT-PCR kit showed excellent sensitivity detecting all positive samples including the weak positive samples (see the table BVD 2011 ring trial results).

Specificity and sensitivity of virotype BVDV RT-PCR Kit

Sample

Reference assay

virotype BVDV RT-PCR Kit result

n

Positive

Negative

Positive

Negative

False positive

False negative

5056

486

4570

484

4568

2

2

Using a mixture of sample types, the virotype BVDV RT-PCR Kit has with this sample set a diagnostic specificity of 99.95% and a diagnostic sensitivity of 99.6%.

BVD 2011 ring trial results

Sample origin

Expected result

virotype BDVD RT-PCR Kit CT

virotype BDVD RT-PCR Kit result

Serum/PI, BVDV-1

Positive

22.71

Positive

Serum/TI, BVDV-1

Positive

28.80

Positive

Serum diagnostic gap, BVDV-1

Positive

30.37

Positive

Serum/PI, BVDV-1

Positive

23.45

Positive

Serum/TI, BVDV-1

Positive

30.30

Positive

Serum diagnostic gap, BVDV-1

Positive

30.94

Positive

Serum/PI, BVDV-1

Positive

26.21

Positive

Serum/PI, BVDV-1

Positive

24.92

Positive

Serum/fetal calf, "HoBi"

Positive

31.43

Positive

Milk/PI

Positive

18.71

Positive

Milk/PI Milk 1:100 diluted

Positive

30.45

Positive

Ear tissue/PI (Caisley tube)

Positive

27.63

Positive

Ear tissue/PI (Caisley tube)

Positive

25.08

Positive

Ear tissue/PI (Caisley tube)

Positive

29.28

Positive

Ear tissue/PI (Allflex tube)

Positive

24.44

Positive

Ear tissue/PI (Allflex tube)

Positive

25.25

Positive

Ear tissue (Caisley tube)

Positive

23.42

Positive

In an independent study conducted by the Friedrich-Loeffler-Institut, the virotype BVDV RT-PCR Kit showed excellent sensitivity detecting all positive samples including the weak positive samples.

Principle

This is a real-time RT-PCR kit for the detection of RNA from bovine viral diarrhea virus. RT-PCR is based on the amplification of specific regions of the pathogen genome. It is a sensitive and specific tool for pathogen detection.

Real-time RT-PCR involves the reverse transcription of viral RNA and subsequent amplification of the cDNA by PCR. The amplified product is detected using fluorescent dyes. These are usually linked to oligonucleotide probes that bind specifically to the amplified product. Monitoring the fluorescence intensities during the PCR run (i.e., in real time) allows the detection of the accumulating product without the need to re-open the reaction tubes afterward.

Procedure

The kit contains a ready-to-use PCR mix and enzyme mix and the required positive and negative controls. With this kit, both reverse transcription and PCR are performed in one reaction tube, reducing the risk of contamination.

RNA is extracted from the sample material (blood, plasma, serum, ear tissue, or milk) using a suitable sample preparation kit. The virotype Tissue Lysis Reagent provides a fast, easy-to-use, and convenient method for RNA preparation from ear tissue samples. The extracted RNA is used for the RT-PCR, which is run on a thermal cycler.

Applications

The virotype BVDV RT-PCR Kit is for detection of RNA from bovine viral diarrhea virus (BVDV) in samples from cattle.